timosaponin-aiii and sarsasapogenin

The rhizome of Anemarrhena asphodeloides (AA, family Liliaceae), which contains furostanol and spirostanol saponins, is a typical herbal medicine that improves learning and memory in rats and inhibits inflammation. In a preliminary study, timosaponin AIII, one of AA main constituents, was metabolized to sarsasapogenin by gut microbiota and inhibited NF-κB activation in lipopolysaccharide (LPS)-stimulated macrophages. Here we have investigated the anti-inflammatory effects of AIII and sarsasapogenin in vitro and in vivo. Both AIII and sarsasapogenin potently inhibited NF-κB and MAPK activation, as well as IRAK1, TAK1, and IκBα phosphorylation in LPS-stimulated macrophages. Further, AIII and sarsasapogenin inhibited the binding of LPS to macrophage Toll-like receptor 4, as well as polarization of M2 to M1 macrophages. Oral administration of AIII and sarsasapogenin inhibited 2,3,4-trinitrobenzene sulfonic acid (TNBS)-induced colon shortening and myeloperoxidase activity in mice, along with reducing NF-κB activation and interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and IL-6 levels, while simultaneously increasing IL-10. Both compounds inhibited Th17 cell differentiation in colonic lamina propria, but induced Treg cell differentiation. Further, AIII and sarsasapogenin inhibited the differentiation of splenic CD4(+) T cells into Th17 cells in vitro. The vitro and in vivo anti-inflammatory effects of sarsasapogenin were more potent than AIII. These results suggest that orally administered AIII may be metabolized to sarsasapogenin by gut microbiota, which may ameliorate inflammatory diseases such as colitis by inhibiting TLR4-NF-κB/MAPK signaling pathway and restoring Th17/Treg cell balance.

Topics: Animals; Anti-Inflammatory Agents; Cell Differentiation; Cells, Cultured; Colitis; Colon; Lipopolysaccharides; Macrophages, Peritoneal; Male; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; NF-kappa B; Peroxidase; Saponins; Spirostans; Steroids; T-Lymphocytes, Regulatory; Th17 Cells; Trinitrobenzenesulfonic Acid

tive: To compare and analyze the quality of Anemarrhena asphodeloides rhizome from different habitats.. Simultaneous determination of nine components in Anemarrhena asphodeloides rhizome by UPLC-TQ/MS was performed on a Phenomenex Kinetex XB-C18 (100 mm x 2.1 mm, 1.7 μm) column with the mobile phase consisted of 0.1% formic acid-acetonitrile (gradient elution) at the flow rate of 0.4 mL/min and thecolumn temperature at 35 degrees C. Multiple reaction mode detection (MRM) in mode was used in this assay.. Nine components were separated totally within 15 min. Good correlation were found between the investigated compounds concentrations and their peak areas within the test ranges with the correlation coefficient from 0.9917 to 0.9992. The average recoveries were from 98.1% to 103.7%, and the RSD of precision was in the range of 1.7% - 4.7%. 0.074-3.620 mg/g for sarsasapogenin, 0.042-2.530 mg/g for timosaponin A III, 22.1- 50.4 mg/g for timosaponon B II, 0.10 -8.28 mg/g for officinalisinin II, 0.64 -7.29 mg/g for anemarsaponin B III, 3.28 -27.40 mg/g for mangiferin, 1.83 - 7.21 mg/g for isomangiferin, 0.36 -9.25 mg/g for neomangiferin and 4.72 x 10(-5) - 1.38 x 10(-3) mg/g for baohuoside I in Anemarrhena asphodeloides rhizome from different habitats were detected.. The method is rapid, accurate and can be used for quality evaluation of Anemarrhena asphodeloides rhizome. The quality of Anemarrhena asphodeloides rhizome from different habitats are different. The saponins content of Anemarrhena asphodeloides rhizome in Hebei is higher than that of the others.

Topics: Anemarrhena; Drugs, Chinese Herbal; Ecosystem; Glucosides; Plant Extracts; Plants, Medicinal; Rhizome; Saponins; Spirostans; Steroids; Triterpenes; Xanthones

To investigate the antiallergic effect of the rhizome of Anemarrhena asphodeloides (AA, family Liliaceae), which was found to inhibit the mouse passive cutaneous anaphylaxis (PCA) reaction induced by the antigen-immunoglobulin E (IgE) complex in preliminary experiments, main steroidal saponins, timosaponins AIII, BIII, and D, were isolated and their inhibitory effects against PCA reaction and scratching behaviors investigated in mice. Oral administration of three main steroidal sapogenins blocked the PCA reaction and scratching behaviors, timosaponin AIII was the most potent. However, intraperitoneal administration of timosaponin AIII showed weak inhibition. To understand its metabolism and antiallergic mechanism, timosaponin AIII was anaerobically incubated with human intestinal microflora to afford a main metabolite, sarsasapogenin. Intraperitoneal administration of sarsasapogenin inhibited allergic reaction more potently than timosaponin AIII. In addition, sarsasapogenin more potently inhibited degranulation and IL-4 protein expression of RBL-2H3 cells induced by IgE-antigen complex than timosaponin AIII. On the basis of these findings, antiallergic effect of AA may be due to those of its steroidal constituents, and that of timosaponin AIII may be activated by using intestinal microflora.

Topics: Anemarrhena; Animals; Anti-Allergic Agents; Antigen-Antibody Complex; Cells, Cultured; Humans; Immunoglobulin E; Interleukin-4; Intestines; Male; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; Passive Cutaneous Anaphylaxis; Pruritus; Rats; Rats, Sprague-Dawley; Rhizome; Saponins; Spirostans; Steroids; Tumor Necrosis Factor-alpha